动物造模 z-bio 2021-11-02 354

　　Objective To analyze the cDNA sequence homology of Cystatin C (CST3) in Mongolian gerbils and establish a prokaryotic expression system for CST3 protein to lay the foundation for the preparation of CST3 antibodies and subsequent gene function studies in Mongolian gerbils.

　　Methods Cloning, homology analysis and codon optimization were performed on the Cst3 cDNA sequence of Mongolian gerbils; the optimized sequence was digested and connected to the pET28a vector to complete the construction of the recombinant CST3 protein expression vector; the vector was transformed into competent cells, Prokaryotic expression of CST3 protein was induced by isopropyl thiogalactoside (IPTG), and verified by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.

　　Result The sequence homology of the Cst3 gene of gerbils and human and mouse Cst3 genes is low. The codon-optimized Cst3 expression sequence of Mongolian gerbils was inserted into the pET28a plasmid, and the CST3 protein expression vector was obtained. After induction with I mmol/L IPTG at 37°C for 12 hours, a large amount of CST3 protein can be obtained.

　　Conclusion The in vitro expression system of the CST3 protein of Mongolian gerbils was successfully constructed.